Peptide nucleic acid based guanidinium compounds

ABSTRACT

Disclosed herein are transmembrane transporter compounds containing guanidinium groups to enhance transport of a polymer backbone across biomembranes. Therapeutic and other biologically active moieties may be attached to the compounds. The polymer backbone may include peptide nucleic acid monomer units.

RELATED APPLICATION

This application claims the benefit of U.S. Provisional Application No.60/720,067, filed Sep. 23, 2005, which is incorporated herein byreference in its entirety.

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates to the fields of chemistry and medicine.More particularly, the present invention relates to guanidiniumcontaining compounds that exhibit enhanced transmembrane transport ofbiologically active molecules.

2. Description of the Related Art

Protein transduction domains (PTD) are short cationic peptides that arecapable of delivering molecules into mammalian cells. They include manydiverse peptide sequences from many diverse sources. The phenomenon thatpeptide sequences can assist molecules across biological barriers isubiquitous in biological systems. Such phenomenon can take place atdifferent levels, including subcellular (e.g., nuclear membranes),cellular (e.g., cell membranes), and tissue levels (e.g., epithelialtissue). Sometimes the same peptide sequences can promote differenttranslocation events at different biological levels.

SUMMARY OF THE INVENTION

One embodiment disclosed herein includes a compound having the formula:

wherein:

each u and each v are separately selected to be an integer from 0 to 10,with the proviso that at least one SPU unit is present between two BBunits;

w is an integer of from 1 to 50;

each L is separately selected to be a linker moiety comprising amolecular fragment having a molecular weight less than about 1000 or Lis absent;

each T is a terminal group separately selected from the group consistingof hydrogen, hydroxy, an amine group, a carboxylic group, a N-terminalpeptide or group that forms an N-terminal peptide bond with BB or SPU, aC-terminal peptide or group that forms a C-terminal peptide bond with BBor SPU, a reporting moiety, a targeting moiety, and a therapeuticmoiety, or each T may be separately absent;

each BB is separately selected to be a backbone moiety comprising anorganic fragment having a molecular weight less than about 5000;

each SPU is separately selected to be a spacer-unit moiety selected fromthe group consisting of mono-substituted, poly-substituted orunsubstituted, straight or branched chain variants of the followingresidues: C₂₋₂₀ alkyl, C₂₋₂₀ heteroalkyl, C₂-C₂₀ alkenyl, C₂-C₂₀alkynyl, C₃-C₂₀ heteroalkenyl, C₃-C₂₄ heteroalkynyl, C₅₋₃₀ aryl, andC₅₋₃₀ heteroaryl;

each R is separately selected from the group consisting of H, anoptionally substituted amino acid, optionally substituted C₃-C₈ aryl,optionally substituted C₂-C₈ heteroaryl, optionally substituted C₁-C₂₀alkyl, optionally substituted C₂-C₂₀ alkenyl, optionally substitutedC₂-C₂₀ alkynyl, optionally substituted C₂-C₈ spirocycloalkylene,optionally substituted C₂-C₈ heterocyclyl, optionally substitutedpyrimidine, and optionally substituted purine, wherein the pyrimidineand purine are optionally linked to the compound via a linker comprisingan optionally substituted C₁-C₈ alkyl, optionally substituted C₂-C₈alkenyl, or optionally substituted C₂-C₈ alkynyl;

each G is separately selected to be a guanidinium group covalentlyattached to a BB group and having the formula:

wherein the point of attachment of G to BB is through a nitrogen atom orthrough R² or each G is separately selected from the group consistingof:

each R¹, R², R³, and R⁴ are separately selected from the groupconsisting of hydrogen, optionally substituted C₁-C₁₂ alkyl, optionallysubstituted C₂-C₁₂ alkenyl, optionally substituted C₂-C₁₂ alkynyl, andpentamethylchroman-6-sulfonyl;

each R¹ is optionally bound to the R², R³, or R⁴ on the same guanidiniumgroup to form a heterocyclic ring;

each R² is optionally bound to the R³ or R⁴ on the same guanidiniumgroup to form a heterocyclic ring; and

-   -   each R³ is optionally bound to the R⁴ on the same guanidinium        group to form a heterocyclic ring.

In some embodiments, each BB-G is separately selected from the groupconsisting of:

In some embodiments, SPU-R has the formula:

In some embodiments, each G is separately selected from the groupconsisting of:

In some embodiments, R¹, R², R³, and R⁴ are hydrogen. In someembodiments, each m, n, p, and q are independently integers from 0 to 1.

In some embodiments, L is a cleavable linker. In some embodiments, thecleavable linker comprises an ester group. In some embodiments, thecleavable linker comprises a disulfide group. In some embodiments, thecleavable linker comprises a first cleavable group and a secondcleavable group, wherein when the first cleavable group is cleaved, thefirst cleavable group is converted to a nucleophilic moiety that isadapted to react with the second cleavable group. In some embodiments,the first group is an amide group and the second group is an estergroup, and cleavage of the amide group yields a free amino group thatreacts with the ester group. In some embodiments, the first group is aphosphate ester group and the second group is a carboxylate ester group,and cleavage of the phosphate ester group yields a free hydroxyl groupthat reacts with the carboxylate ester group.

In some embodiments, each L is separately selected from the groupconsisting of: an oligopeptide comprising 1 to 12 amino acid residues,an optionally substituted C₁-C₁₂ alkyl, an optionally substituted C₂-C₁₂alkenyl, an optionally substituted C₂-C₁₂ alkynyl, and an optionallysubstituted C₃₋₁₂ cyclic alkyl, alkenyl, alkynyl, or aromatic moiety.

In some embodiments, each L is separately selected from the groupconsisting of: —C(═O)NH—, —C(═O)NHO—, —C(═O)NHNH—, —S(═O)(═O)NR′—,—P(═O)(—OR′)NR″—, —SS—, —CH₂NR′—, —CH₂NR′—, —CH₂C(═O)NR′—, —C(═O)O—,—C(═S)NR′—, —S(═O)(═O)CH₂—, —SOCH₂— and —OC(═O)NR′—, wherein each R′ andR″ are separately selected from the group consisting of hydrogen,optionally substituted C₁-C₁₂ alkyl, optionally substituted C₂-C₁₂alkenyl, and optionally substituted C₂-C₁₂ alkynyl.

In some embodiments, L is absent.

In various embodiments, at least one T comprises a polypeptide, proteinantigen, tumor antigen, tisane moiety, antimicrobial agent, metal ion,or cleavable linker, including cleavable linkers having an ester groupor a disulfide group such as described above. In some embodiments, atleast one T comprises a peptide nucleic acid.

In some embodiments, each R is separately selected from the groupconsisting of an optionally substituted pyrimidine and an optionallysubstituted purine, the pyrimidine and purine optionally linked to thecompound via a linker comprising an optionally substituted C₁-C₈ alkyl,optionally substituted C₂-C₈ alkenyl, or optionally substituted C₂-C₈alkynyl. In some embodiments, each R is separately selected from thegroup consisting of uracil, thymine, cytosine, adenine, and guanine, theuracil, thymine, cytosine, adenine, and guanine optionally linked to thecompound via a linker comprising an optionally substituted C₁-C₈ alkyl,optionally substituted C₂-C₈ alkenyl, or optionally substituted C₂-C₈alkynyl. In some embodiments, at least one R has the formula:

Another embodiment disclosed herein includes a compound as describedabove having the formula:

wherein each m, n, p, and q are independently integers from 0 to 6 and ris an integer from 1 to 10.

One embodiment includes a compound having the formula:

In some embodiments, at least one R has the formula:

One embodiment includes a compound having the formula:

Another embodiment disclosed herein includes a polymer, comprising atleast two arginine subunits and at least two peptide nucleic acidsubunits. In some embodiments, at least two arginine subunits areadjacent to each other. In other embodiments, no arginine subunits areadjacent to each other. In some embodiments, the peptide nucleic acidsubunit has the formula:

wherein R is selected from the group consisting of an optionallysubstituted pyrimidine and an optionally substituted purine, wherein thepyrimidine and purine are optionally linked to the compound via a linkercomprising an optionally substituted C₁-C₈ alkyl, optionally substitutedC₂-C₈ alkenyl, or optionally substituted C₂-C₈ alkynyl.

Another embodiment disclosed herein includes a polymer, comprising atleast two arginine subunits and at least two spacer subunits. In someembodiments, at least two arginine subunits are adjacent to each other.In some embodiments, no arginine subunits are adjacent to each other. Insome embodiments, the spacer subunit has the formula:

wherein R is selected from the group consisting of H, an optionallysubstituted amino acid, optionally substituted C₃-C₂₀ aryl, optionallysubstituted C₂-C₂₀ heteroaryl, optionally substituted C₁-C₂₀ alkyl,optionally substituted C₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀alkynyl, optionally substituted C₂-C₈ spirocycloalkylene, optionallysubstituted C₂-C₈ heterocyclyl, optionally substituted pyrimidine, andoptionally substituted purine, wherein the pyrimidine and purine areoptionally linked to the compound via a linker comprising an optionallysubstituted C₁-C₈ alkyl, optionally substituted C₂-C₈ alkenyl, oroptionally substituted C₂-C₈ alkynyl.

Another embodiment disclosed herein includes a method for enhancingtransport of a biologically active moiety across a biological membrane,comprising contacting a biological membrane with a compound of claim 1,wherein at least one T comprises a biologically active moiety, wherebythe contacting is effective to promote transport of the compound acrossthe biological membrane at a rate that is greater than a trans-membranetransport rate of the biologically active moiety in non-conjugated form.In some embodiments, the biological membrane is a eukaryotic cellmembrane. In some embodiments, the eukaryotic cells selected from thegroup consisting of mammalian cells, cancer cells, insect cells, plantcells, and yeast cells. In some embodiments, the biological membrane isan epithelial layer in a body. In some embodiments, the epithelial layeris selected from the group consisting of skin, mucosmembrane, and brainblood barriers. In some embodiments, the biological membrane is aprokaryotic cell membrane.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 depicts a fluorescence micrograph of cells exposed to a peptidenucleic acid transporter molecule and a control fluorescence micrograph.

FIG. 2 depicts a fluorescence micrograph of skin tissue exposed to apeptide nucleic acid transporter molecule and a control fluorescencemicrograph.

DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT

Peptide Nucleic Acid (PNA), a synthetic analogue of DNA and RNA in whichthe natural sugar-phosphate backbone has been replaced byN-(2-aminoethyl)glycine exhibits high thermal stability and stability toboth nucleases and proteases. Thus, PNAs may be suitable as ruggedbackbones for therapeutic delivery. However, PNAs are not readily takenup by mammalian cells and have a tendency to aggregate when in watersolution. Accordingly, in one embodiment, transmembrane transport of PNAand other backbone containing molecules are enhanced by the attachmentof transport promoting moieties. Oligoarginine sequences generated byreplacing non-arginine residues with arginine in the HIV Tat 47-57segment have been demonstrated to be effective at assisting molecules tocross cellular membranes and mammalian skin tissues. Thus, in oneembodiment, guanidinium and related moieties are attached to PNA orother backbones to promote transmembrane transport.

In some embodiments, a guanidinium containing transporter molecule isprovided having one or more therapeutic moieties attached thereto. Thus,the transporter-therapeutic complex can be readily transported into acell where the therapeutic moiety can have therapeutic effect. Anytherapeutic moiety that can be covalently bound to the transportermolecules disclosed herein may be used. Non-limiting examples of sometherapeutic moieties include: immunosuppressives, antibacterials,antifungals, antivirals, antiproliferatives, hormones,antiinflammatories, vitamins, analgesics, diagnostics and imagingcontrast agents. Some specific examples include: conjugatedglucocorticoids for treating inflammatory skin diseases, conjugatedretinoids for treating acne, skin cancer and psoriasis, conjugatedcytotoxic and immunosuppressive drugs for treating cancer and leukemia,conjugated antiinflammatory and antifungal agents, conjugatedantihistamines, conjugated anagelsics, conjugated photochemotherapeuticagents, sunscreen, conjugated antibiotics for treating bacteriainfections, conjugated anti-neoplastic agents for treating cancers,conjugated antiinflammatory agents, bronchodialators andimmunosuppressive drugs for treating pulmonary conditions, conjugatedneurotransmitters, and analgesics and CNS drugs that are capable ofcrossing the blood brain barrier.

In some embodiments, the therapeutic moiety is attached to thetransporter molecule via a cleavable linker moiety. The cleavable linkermoiety may be such that once the transporter-therapeutic complex istransported into a cell, the linker is cleaved, freeing the therapeuticmoiety from the complex. Those of skill in the art will recognize manycleavable linkers suitable for this purpose. Non-limiting examples ofcleavable linkers include moieties containing di-sulfide functionalitiesor acid-degradable functionalities such as esters. In some embodiments,a targeting moiety may also be attached to the multivalent argininetransporter molecule. The targeting moiety may be any moiety thatpreferentially binds to receptors on one or more cell types. Thus, atransporter-targeting-therapeutic complex is provided thatpreferentially is transported into one or more cell types where thetherapeutic moiety can have therapeutic effect. Those of skill in theart will recognize many suitable targeting moieties. Non-limitingexamples of some targeting moieties include: small molecules, oligopeptides, oligonucleotides, and macromolecules obtainable from syntheticor natural resources that have specific interactions with moleculartargets implicated in the following diseases, including Acutelymphoblastic leukemia, Advanced pancreatic tumor, Affective disorder,AIDS, Allergic rhinitis, Allergy, Alzheimer's, Analgesic, Anesthesia,ANF degradation, Angiogenesis, Anxiety, Arthritis, Asthma, Autoimmunedisease, Bacterial infection, Baldness, Blood coagulation, Bone Loss,Brain ischaemia, Breast cancer, Calcium deficiency, Carcinoid syndrome,Cardiac failure, Cardiovascular disease, Chronic myelogenous leukemia,Cognitive dysfunction, Colon cancer, Common cold, Common roundworm,Congestive heart failure, Cystic fibrosis, Dementia, Depression,Diabetes, Diabetic retinopathy, Diarrhea, Drug dependence, Erectiledysfunction, Fever, Fungal infection, Gastric tumor, Glaucoma, Gout,Heart disease, Heart failure, Helminth infection, Hepatitis C, Herpes,High blood glucose level, High blood sugar level, High cholesterol,Hirsutism, Hormone-dependent tumors, Human African trypanosomiasis,Hypertension, Hyperthyroidism, Hypocalcaemia, Immune response,Immunodeficiency, Inflammation, Influenza A and B, Insomnia, Irritablebowel syndrome, Kidney failure, Leukemia, Liposarcoma, Liver, Localanesthetic, Lung cancer, Lupus, Malaria, Malignant pain, Melanoma,Metastasis, Migraine, Morning sickness, Motion sickness, Motor disorder,Movement disorder, Nasal congestion, Neurodegeneration, Neuropathic,Obesity, Obstructive pulmonary disease, Ocular hypertension/glaucoma,Osteoporosis, Ovarian, Pain, Parkinson's, Peptic ulcer,Phaeochromocytoma, Platelet adhesion, Platelet disease, Posteriorpituitary disorder, Postsurgical pain, Prostate adenocarcinoma, Prostatetumor, Prostatic hyperplasia, Psychiatric illness, Psychomotor,Reproduction diseases, Respiration diseases, Rheumatoid, Riboflavindeficiency, Schizophrenia, Seizure, Smoking addiction, Solid tumor,Thiamine deficiency, Tuberculosis, Urinary tract infection, Urticaria,Uterus contraction, Vascular disease, Viral infection, Visceral, VitaminA deficiency, Vitamin B12 deficiency, Vitamin B6 deficiency, Vitamin Cdeficiency, Vitamin D deficiency, Vomiting, and Zollinger-Ellisonsyndrome.

In one embodiment, the transporter molecules for use as described hereinhave the structure:

BB represents a back-bone moiety to which guanidinium moieties G may beattached. SPU represents a spacer-unit moiety to provide molecularspacers between multiple BB-G units. In some embodiments, SPU-R mayinclude PNA moieties. Thus, the SPU-R units together with the BB unitsmay provide a backbone structure that includes PNA units and attachedguanidinium moieties G. Accordingly, a rugged PNA containing moleculemay be provided that also has transmembrane functionality due toattached guanidinium moieties. Because a given molecule may havemultiple SPU and/or BB units, each separate SPU and BB unit may beselected separately. Thus, some or all of the SPU units may be differentfrom other SPU units and some or all of the BB units may be differentfrom other BB units.

Each u and each v may be separately selected to be an integer from 0 to10. Thus, each repeated unit w may contain multiple SPU and BB units oroptionally may contain no SPU unit or no BB unit. Accordingly, in somecases, the transporter molecule may include domains of multiple BB unitsbound together in a chain with no intervening SPU unit. Similarly, thetransporter molecule may contain domains of multiple SPU units boundtogether in a chain with no intervening BB units. w may be an integer offrom 1 to 50. As noted above, each SPU-R unit and each BB-G unit,whether within the same w unit or different w units may be differentfrom other SPU-R and BB-G units in the molecule.

BB is a backbone moiety comprising an organic fragment that provides ascaffold upon which guanidinium moieties G may be attached. In onesimple example, BB-G is an arginine residue. In one embodiment, BB is anorganic fragment having a molecular weight less than about 10,000. Inone embodiment, BB is an organic fragment having a molecular weight lessthan about 5,000. In one embodiment, BB is an organic fragment having amolecular weight less than about 1,000.

Some non-limiting examples of backbone moieties BB with attachedguanidinium groups G include:

G represents a guanidinium moiety and may have the structure:

where the point of attachment of G to BB is through the nitrogen atom orthrough R² or, alternatively, each G may be selected from the groupconsisting of:

Each R¹, R², R³, and R⁴ may be separately selected from the groupconsisting of hydrogen, optionally substituted C₁-C₁₂ alkyl, optionallysubstituted C₂-C₁₂ alkenyl, optionally substituted C₂-C₁₂ alkynyl, andan N protecting group such as pentamethylchroman-6-sulfonyl. The wavyline indicates where guanidinium moiety G attaches to a backbone moietyBB. R¹, R², R³, and R⁴ on each guanidinium moiety G may be the same ordifferent from R¹, R², R³, and R⁴ groups on other guanidinium moietiesG. In some embodiments, R¹ may be optionally bound to a R², R³, or R⁴group on the same guanidinium group to form a heterocyclic ring withinthe guanidinium group. Similarly, in some embodiments, R² may beoptionally bound to a R³ or R⁴ group on the same guanidinium group toform a heterocyclic ring or R³ may be optionally bound to the R⁴ on thesame guanidinium group to form a heterocyclic ring.

Some non-limiting examples of G include:

Each SPU is a spacer unit separately selected from the group consistingof mono-substituted, poly-substituted or unsubstituted, straight orbranched chain variants of the following residues: C₂₋₂₀ alkyl, C₂₋₂₀heteroalkyl, C₂-C₂₀ alkenyl, C₂-C₂₀ alkynyl, C₃-C₂₀ heteroalkenyl,C₃-C₂₄ heteroalkynyl, C₅₋₃₀ aryl, and C₅₋₃₀ heteroaryl. In anon-limiting example, an SPU-R has the structure of:

where the wavy lines indicate the attachment points of SPU to the restof the molecule.

Each R bound to an SPU unit may be separately selected from the groupconsisting of H, optionally substituted amino acid, optionallysubstituted C₃-C₈ aryl, optionally substituted C₂-C₈ heteroaryl,optionally substituted C₁-C₂₀ alkyl, optionally substituted C₂-C₂₀alkenyl, optionally substituted C₂-C₂₀ alkynyl, optionally substitutedC₂-C₈ spirocycloalkylene, optionally substituted C₂-C₈ heterocyclyl,optionally substituted pyrimidine, and optionally substituted purine.When pyrimidine and purine moieties are present, they may be optionallylinked to the SPU unit via a linker comprising an optionally substitutedC₁-C₈ alkyl, optionally substituted C₂-C₈ alkenyl, or optionallysubstituted C₂-C₈ alkynyl. In some embodiments, each R is separatelyselected from the group consisting of uracil, thymine, cytosine,adenine, and guanine. The uracil, thymine, cytosine, adenine, andguanine may optionally be linked to the compound via a linker comprisingan optionally substituted C₁-C₈ alkyl, optionally substituted C₂-C₈alkenyl, or optionally substituted C₂-C₈ alkynyl. Some non-limitingexamples of R include:

where the wavy line indicates the point of attachment of R to the SPUunit.

L is a linker moiety comprising a molecular fragment that links T to therest of the molecule. Each separate linker moiety L may be the same ordifferent from other linker moieties L. As discussed above, linkermoiety L may optionally be a cleavable linker designed to cleave aterminal group T from the transporter molecule under certain conditions.The cleavable linker may include an ester group that can by hydrolyzedby an enzyme within a subject. Alternatively, the cleavable linker mayinclude a disulfide group that can also be enzymatically cleaved. Insome embodiments, the cleavable linker may include two cleavable groupssuch that when one cleavable group is cleaved, it is converted to anucleophilic moiety that reacts with the other cleavable group. In oneembodiment, the first cleavable group is an amide group and the secondgroup is an ester group so that cleavage of the amide group yields afree amino group that reacts with the ester group. In anotherembodiment, the first cleavable group is a phosphate ester group and thesecond group is a carboxylate ester group so that cleavage of thephosphate ester group yields a free hydroxyl group that reacts with thecarboxylate ester group. In some embodiments, L is a molecular fragmenthaving a molecular weight less than about 5000. In some embodiments, Lis a molecular fragment having a molecular weight less than bout 1000.In some embodiments, L is a molecular fragment having a molecular weightless than about 500.

T represents terminal groups attached to each end of the molecule.Terminal group T may serve the function of merely terminating the chainor may also include therapeutic, targeting, or reporting moieties. Whena terminal group T includes a therapeutic moiety, the moiety may belinked to the rest of the molecule through a cleavable linker such asdescribed above. Each T group may be the same or different from theother T group. In some embodiments, each terminal group T may beseparately selected from the group consisting of hydrogen, hydroxy, anamine group, a carboxylic group, an N-terminal peptide or group thatforms an N-terminal peptide bond with BB or SPU, a C-terminal peptide orgroup that forms a C-terminal peptide bond with BB or SPU, a reportingmoiety, a therapeutic moiety, and a targeting moiety. Alternatively, a Tgroup may be absent if a terminal BB group does not require aterminating group, such as when a guanidinium moiety G terminates thebackbone chain.

Non-limiting examples of T include a polypeptide, a protein antigen, atumor antigen, a tisane moiety, a metal ion, or an antimicrobial agent.

In some embodiments, the compound described above has the formula:

Each m, n, p, and q may be separately selected to be integers from 0 to6. Accordingly, in some cases, the transporter molecule may includedomains of multiple G containing moieties bound together in a chain withno intervening R containing moieties. Similarly, the transportermolecule may contain domains of multiple R containing moieties boundtogether in a chain with no intervening G containing moieties. r may bean integer from 1 to 10. Each G and each R, whether within the same runit or different r units may be different from other G and R groups inthe molecule. G and R are as described above. In one embodiment, each m,n, p, and q are separately integers from 0 to 1.

Some non-limiting examples of transporter molecules for use as describedherein include the following molecules:

Some embodiments include a polymer that has at least two argininesubunits and at least two peptide nucleic acid subunits. In oneembodiment, at least two arginine subunits are adjacent to each other.In another embodiment, no arginine subunits are adjacent to each other.In some embodiments, the peptide nucleic acid subunit has the formula:

where R is selected from the group consisting of an optionallysubstituted pyrimidine and an optionally substituted purine, wherein thepyrimidine and purine are optionally linked to the compound via a linkercomprising an optionally substituted C₁-C₈ alkyl, optionally substitutedC₂-C₈ alkenyl, or optionally substituted C₂-C₈ alkynyl.

Other embodiments include a polymer having at least two argininesubunits and at least two spacer subunits. In some embodiments, thespacer unit has the formula:

where R is selected from the group consisting of H, an optionallysubstituted amino acid, optionally substituted C₃-C₂₀ aryl, optionallysubstituted C₂-C₂₀ heteroaryl, optionally substituted C₁-C₂₀ alkyl,optionally substituted C₂-C₂₀ alkenyl, optionally substituted C₂-C₂₀alkynyl, optionally substituted C₂-C₈ spirocycloalkylene, optionallysubstituted C₂-C₈ heterocyclyl, optionally substituted pyrimidine, andoptionally substituted purine, wherein the pyrimidine and purine areoptionally linked to the compound via a linker comprising an optionallysubstituted C₁-C₈ alkyl, optionally substituted C₂-C₈ alkenyl, oroptionally substituted C₂-C₈ alkynyl.

Where the compounds disclosed herein have at least one chiral center,they may exist as a racemate or as enantiomers. It should be noted thatall such isomers and mixtures thereof are included in the scope of thepresent invention. Furthermore, some of the crystalline forms for thecompounds of disclosed herein may exist as polymorphs. Such polymorphsare included in one embodiment of the present invention. In addition,some of the compounds of the present invention may form solvates withwater (i.e., hydrates) or common organic solvents. Such solvates areincluded in one embodiment of the present invention.

In some embodiments, the compounds disclosed herein may be provided in apharmaceutically acceptable salt. The term “pharmaceutically acceptablesalt” refers to a salt of a compound that does not cause significantirritation to an organism to which it is administered and does notabrogate the biological activity and properties of the compound. In someembodiments, the salt is an acid addition salt of the compound.Pharmaceutical salts can be obtained by reacting a compound withinorganic acids such as hydrohalic acid (e.g., hydrochloric acid orhydrobromic acid), sulfuric acid, nitric acid, phosphoric acid and thelike. Pharmaceutical salts can also be obtained by reacting a compoundwith an organic acid such as aliphatic or aromatic carboxylic orsulfonic acids, for example acetic, succinic, lactic, malic, tartaric,citric, ascorbic, nicotinic, methanesulfonic, ethanesulfonic,p-toluensulfonic, salicylic or naphthalenesulfonic acid. Pharmaceuticalsalts can also be obtained by reacting a compound with a base to form asalt such as an ammonium salt, an alkali metal salt, such as a sodium ora potassium salt, an alkaline earth metal salt, such as a calcium or amagnesium salt, a salt of organic bases such as dicyclohexylamine,N-methyl-D-glucamine, tris(hydroxymethyl)methylamine, C₁-C₇ alkylamine,cyclohexylamine, triethanolamine, ethylenediamine, and salts with aminoacids such as arginine, lysine, and the like.

If the manufacture of pharmaceutical formulations involves intimatemixing of the pharmaceutical excipients and the active ingredient in itssalt form, then it may be desirable to use pharmaceutical excipientswhich are non-basic, that is, either acidic or neutral excipients.

In various embodiments, the compounds disclosed herein can be usedalone, in combination with other compounds disclosed herein, or incombination with one or more other agents that are therapeuticallyactive.

The term “halogen atom,” as used herein, means any one of theradio-stable atoms of column 7 of the Periodic Table of the Elements,e.g., fluorine, chlorine, bromine, or iodine, with fluorine and chlorinebeing preferred.

The term “ester” refers to a chemical moiety with formula—(R)_(n)—COOR′, where R and R′ are independently selected from the groupconsisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded through a ringcarbon) and heteroalicyclic (bonded through a ring carbon), and where nis 0 or 1.

An “amide” is a chemical moiety with formula —(R)_(n)—C(O)NHR′ or—(R)_(n)—NHC(O)R′, where R and R′ are independently selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded througha ring carbon) and heteroalicyclic (bonded through a ring carbon), andwhere n is 0 or 1. An amide may be an amino acid or a peptide moleculeattached to a molecule of the present invention, thereby forming aprodrug.

Any amine, hydroxy, or carboxyl side chain on the compounds of thepresent invention can be esterified or amidified. The procedures andspecific groups to be used to achieve this end are known to those ofskill in the art and can readily be found in reference sources such asGreene and Wuts, Protective Groups in Organic Synthesis, 3^(rd) Ed.,John Wiley & Sons, New York, N.Y., 1999, which is incorporated herein inits entirety.

The term “aromatic” refers to an aromatic group which has at least onering having a conjugated pi electron system and includes bothcarbocyclic aryl (e.g., phenyl) and heterocyclic aryl groups (e.g.,pyridine). The term includes monocyclic or fused-ring polycyclic (i.e.,rings which share adjacent pairs of carbon atoms) groups. The term“carbocyclic” refers to a compound which contains one or more covalentlyclosed ring structures, and that the atoms forming the backbone of thering are all carbon atoms. The term thus distinguishes carbocyclic fromheterocyclic rings in which the ring backbone contains at least one atomwhich is different from carbon. The term “heteroaromatic” refers to anaromatic group which contains at least one heterocyclic ring.

The term “alkyl,” as used herein, means any unbranched or branched,substituted or unsubstituted, saturated hydrocarbon. The alkyl moiety,may be branched, straight chain, or cyclic. The alkyl group may have 1to 20 carbon atoms (whenever it appears herein, a numerical range suchas “1 to 20” refers to each integer in the given range; e.g., “1 to 20carbon atoms” means that the alkyl group may consist of 1 carbon atom, 2carbon atoms, 3 carbon atoms, etc., up to and including 20 carbon atoms,although the present definition also covers the occurrence of the term“alkyl” where no numerical range is designated). The alkyl group mayalso be a medium size alkyl having 1 to 10 carbon atoms. The alkyl groupcould also be a lower alkyl having 1 to 5 carbon atoms. The alkyl groupmay be designated as “C₁-C₄ alkyl” or similar designations. By way ofexample only, “C₁-C₄ alkyl” indicates that there are one to four carbonatoms in the alkyl chain, i.e., the alkyl chain is selected from thegroup consisting of methyl, ethyl, propyl, iso-propyl, n-butyl,iso-butyl, sec-butyl, and t-butyl.

The alkyl group may be substituted or unsubstituted. When substituted,the substituent group(s) is (are) one or more group(s) individually andindependently selected from substituted or unsubstituted cycloalkyl,substituted or unsubstituted cylcloalkenyl, substituted or unsubstitutedaryl, substituted or unsubstituted heteroaryl, substituted orunsubstituted heteroaryloxy, heterocyclyl, heterocyclooxy,heteroalicyclyl, hydroxy, substituted or unsubstituted alkoxy,substituted or unsubstituted aryloxy, acyl, thiol, substituted orunsubstituted thioalkoxy, alkylthio, arylthio, cyano, halo, carbonyl,thiocarbonyl, acylalkyl, acylamino, acyloxy, aminoacyl, aminoacyloxy,oxyacylamino, keto, thioketo, O-carbamyl, N-carbamyl, O-thiocarbamyl,N-thiocarbamyl, C-amido, N-amido, S-sulfonamido, N-sulfonamido,C-carboxy, O-carboxy, isocyanato, thiocyanato, isothiocyanato, nitro,silyl, trihalomethanesulfonyl, and substituted or unsubstituted amino,including mono- and di-substituted amino groups, and the protectedderivatives thereof, hydroxyamino, alkoxyamino, nitro, —SO-alkyl,—SO-substituted alkyl, —SO-aryl, —SO-heteroaryl, —SO₂-alkyl,—SO₂-substituted alkyl, —SO₂-aryl and —SO₂-heteroaryl. Typical alkylgroups include, but are in no way limited to, methyl, ethyl, propyl,isopropyl, butyl, isobutyl, tertiary butyl, pentyl, hexyl, ethenyl,propenyl, butenyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, andthe like. Wherever a substituent is described as being “optionallysubstituted” that substituent may be substituted with one of the abovesubstituents.

In the present context, the term “cycloalkyl” is intended to coverthree-, four-, five-, six-, seven-, and eight- or more membered ringscomprising carbon atoms only. A cycloalkyl can optionally contain one ormore unsaturated bonds situated in such a way, however, that an aromaticpi-electron system does not arise. Some examples of “cycloalkyl” are thecarbocycles cyclopropane, cyclobutane, cyclopentane, cyclopentene,cyclopentadiene, cyclohexane, cyclohexene, 1,3-cyclohexadiene,1,4-cyclohexadiene, cycloheptane, or cycloheptene.

An “alkenyl” moiety refers to a group consisting of at least two carbonatoms and at least one carbon-carbon double bond. An alkenyl may beunbranched or branched, substituted or unsubstituted, unsaturatedhydrocarbon including polyunsaturated hydrocarbons. In some embodiments,the alkenyl is a C₁-C₆ unbranched, mono-unsaturated or di-unsaturated,unsubstituted hydrocarbons. The term “cycloalkenyl” refers to anynon-aromatic hydrocarbon ring, preferably having five to twelve atomscomprising the ring.

An “alkyne” moiety refers to a group consisting of at least two carbonatoms and at least one carbon-carbon triple bond.

Unless otherwise indicated, the substituent “R” appearing by itself andwithout a number designation refers to a substituent selected from thegroup consisting of alkyl, cycloalkyl, aryl, heteroaryl (bonded througha ring carbon) and heteroalicyclyl (bonded through a ring carbon).

The term “alkoxy” refers to any unbranched, or branched, substituted orunsubstituted, saturated or unsaturated ether, with C₁-C₆ unbranched,saturated, unsubstituted ethers being preferred, with methoxy beingpreferred, and also with dimethyl, diethyl, methyl-isobutyl, andmethyl-tert-butyl ethers also being preferred. The term “cycloalkoxy”refers to any non-aromatic hydrocarbon ring, preferably having five totwelve atoms comprising the ring.

An “O-carboxy” group refers to a RC(═O)O— group, where R is as definedherein.

A “C-carboxy” group refers to a —C(═O)OR groups where R is as definedherein.

An “acetyl” group refers to a —C(═O)CH₃, group.

A “trihalomethanesulfonyl” group refers to a X₃CS(═O)₂— group where X isa halogen.

A “cyano” group refers to a —CN group.

An “isocyanato” group refers to a —NCO group.

A “thiocyanato” group refers to a —CNS group.

An “isothiocyanato” group refers to a —NCS group.

A “sulfinyl” group refers to a —S(═O)—R group, with R as defined herein.

A “S-sulfonamido” group refers to a —S(═O)₂NR, group, with R as definedherein.

A “N-sulfonamido” group refers to a RS(═O)₂NH— group with R as definedherein.

A “trihalomethanesulfonamido” group refers to a X₃CS(═O)₂NR— group withX and R as defined herein.

An “O-carbamyl” group refers to a —OC(═O)—NR, group-with R as definedherein.

An “N-carbamyl” group refers to a ROC(═O)NH— group, with R as definedherein.

An “O-thiocarbamyl” group refers to a —OC(═S)—NR, group with R asdefined herein.

An “N-thiocarbamyl” group refers to an ROC(═S)NH— group, with R asdefined herein.

A “C-amido” group refers to a —C(═O)—NR₂ group with R as defined herein.

An “N-amido” group refers to a RC(═O)NH— group, with R as definedherein.

The term “perhaloalkyl” refers to an alkyl group where all of thehydrogen atoms are replaced by halogen atoms.

The term “acylalkyl” refers to a RC(═O)R′— group, with R as definedherein, and R′ being a diradical alkylene group. Examples of acylalkyl,without limitation, may include CH₃C(═O)CH₂—, CH₃C(═O)CH₂CH₂—,CH₃CH₂C(═O)CH₂CH₂—, CH₃C(═O)CH₂CH₂CH₂—, and the like.

Unless otherwise indicated, when a substituent is deemed to be“optionally substituted,” it is meant that the substituent is a groupthat may be substituted with one or more group(s) individually andindependently selected from cycloalkyl, aryl, heteroaryl,heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio,arylthio, cyano, halo, carbonyl, thiocarbonyl, O-carbamyl, N-carbamyl,O-thiocarbamyl, N-thiocarbamyl, C-amido, N-amido, S-sulfonamido,N-sulfonamido, C-carboxy, O-carboxy, isocyanato, thiocyanato,isothiocyanato, nitro, silyl, trihalomethanesulfonyl, and amino,including mono- and di-substituted amino groups, and the protectedderivatives thereof. The protecting groups that may form the protectivederivatives of the above substituents are known to those of skill in theart and may be found in references such as Greene and Wuts, above.

The term “heterocyclyl” is intended to mean three-, four-, five-, six-,seven-, and eight- or more membered rings wherein carbon atoms togetherwith from 1 to 3 heteroatoms constitute the ring. A heterocyclyl canoptionally contain one or more unsaturated bonds situated in such a way,however, that an aromatic pi-electron system does not arise. Theheteroatoms are independently selected from oxygen, sulfur, andnitrogen.

A heterocyclyl can further contain one or more carbonyl or thiocarbonylfunctionalities, so as to make the definition include oxo-systems andthio-systems such as lactams, lactones, cyclic imides, cyclicthioimides, cyclic carbamates, and the like.

Heterocyclyl rings can optionally also be fused to aryl rings, such thatthe definition includes bicyclic structures. Typically such fusedheterocyclyl groups share one bond with an optionally substitutedbenzene ring. Examples of benzo-fused heterocyclyl groups include, butare not limited to, benzimidazolidinone, tetrahydroquinoline, andmethylenedioxybenzene ring structures.

Some examples of “heterocyclyls” include, but are not limited to,tetrahydrothiopyran, 4H-pyran, tetrahydropyran, piperidine, 1,3-dioxin,1,3-dioxane, 1,4-dioxin, 1,4-dioxane, piperazine, 1,3-oxathiane,1,4-oxathiin, 1,4-oxathiane, tetrahydro-1,4-thiazine, 2H-1,2-oxazine,maleimide, succinimide, barbituric acid, thiobarbituric acid,dioxopiperazine, hydantoin, dihydrouracil, morpholine, trioxane,hexahydro-1,3,5-triazine, tetrahydrothiophene, tetrahydrofuran,pyrroline, pyrrolidine, pyrrolidone, pyrrolidione, pyrazoline,pyrazolidine, imidazoline, imidazolidine, 1,3-dioxole, 1,3-dioxolane,1,3-dithiole, 1,3-dithiolane, isoxazoline, isoxazolidine, oxazoline,oxazolidine, oxazolidinone, thiazoline, thiazolidine, and1,3-oxathiolane. Binding to the heterocycle can be at the position of aheteroatom or via a carbon atom of the heterocycle, or, for benzo-fusedderivatives, via a carbon of the benzenoid ring.

In the present context the term “aryl” is intended to mean a carbocyclicaromatic ring or ring system. Moreover, the term “aryl” includes fusedring systems wherein at least two aryl rings, or at least one aryl andat least one C₃₋₈-cycloalkyl share at least one chemical bond. Someexamples of “aryl” rings include optionally substituted phenyl,naphthalenyl, phenanthrenyl, anthracenyl, tetralinyl, fluorenyl,indenyl, and indanyl. The term “aryl” relates to aromatic, including,for example, benzenoid groups, connected via one of the ring-formingcarbon atoms, and optionally carrying one or more substituents selectedfrom heterocyclyl, heteroaryl, halo, hydroxy, amino, cyano, nitro,alkylamido, acyl, C₁₋₆ alkoxy, C₁₋₆ alkyl, C₁₋₆ hydroxyalkyl, C₁₋₆aminoalkyl, C₁₋₆ alkylamino, alkylsulfenyl, alkylsulfinyl,alkylsulfonyl, sulfamoyl, or trifluoromethyl. The aryl group can besubstituted at the para and/or meta positions. In other embodiments, thearyl group can be substituted at the ortho position. Representativeexamples of aryl groups include, but are not limited to, phenyl,3-halophenyl, 4-halophenyl, 3-hydroxyphenyl, 4-hydroxyphenyl,3-aminophenyl, 4-aminophenyl, 3-methylphenyl, 4-methylphenyl,3-methoxyphenyl, 4-methoxyphenyl, 4-trifluoromethoxyphenyl3-cyanophenyl, 4-cyanophenyl, dimethylphenyl, naphthyl, hydroxynaphthyl,hydroxymethylphenyl, trifluoromethylphenyl, alkoxyphenyl,4-morpholin-4-ylphenyl, 4-pyrrolidin-1-ylphenyl, 4-pyrazolylphenyl,4-triazolylphenyl, and 4-(2-oxopyrrolidin-1-yl)phenyl.

In the present context, the term “heteroaryl” is intended to mean aheterocyclic aromatic group where one or more carbon atoms in anaromatic ring have been replaced with one or more heteroatoms selectedfrom the group comprising nitrogen, sulfur, phosphorous, and oxygen.

Furthermore, in the present context, the term “heteroaryl” comprisesfused ring systems wherein at least one aryl ring and at least oneheteroaryl ring, at least two heteroaryl rings, at least one heteroarylring and at least one heterocyclyl ring, or at least one heteroaryl ringand at least one cycloalkyl ring share at least one chemical bond.

The term “heteroaryl” is understood to relate to aromatic, C₃₋₈ cyclicgroups further containing one oxygen or sulfur atom or up to fournitrogen atoms, or a combination of one oxygen or sulfur atom with up totwo nitrogen atoms, and their substituted as well as benzo- andpyrido-fused derivatives, for example, connected via one of thering-forming carbon atoms. Heteroaryl groups can carry one or moresubstituents, selected from halo, hydroxy, amino, cyano, nitro,alkylamido, acyl, C₁₋₆-alkoxy, C₁₋₆-alkyl, C₁₋₆-hydroxyalkyl,C₁₋₆-aminoalkyl, C₁₋₆-alkylamino, alkylsulfenyl, alkylsulfinyl,alkylsulfonyl, sulfamoyl, or trifluoromethyl. In some embodiments,heteroaryl groups can be five- and six-membered aromatic heterocyclicsystems carrying 0, 1, or 2 substituents, which can be the same as ordifferent from one another, selected from the list above. Representativeexamples of heteroaryl groups include, but are not limited to,unsubstituted and mono- or di-substituted derivatives of furan,benzofuran, thiophene, benzothiophene, pyrrole, pyridine, indole,oxazole, benzoxazole, isoxazole, benzisoxazole, thiazole, benzothiazole,isothiazole, imidazole, benzimidazole, pyrazole, indazole, tetrazole,quionoline, isoquinoline, pyridazine, pyrimidine, purine and pyrazine,furazan, 1,2,3-oxadiazole, 1,2,3-thiadiazole, 1,2,4-thiadiazole,triazole, benzotriazole, pteridine, phenoxazole, oxadiazole,benzopyrazole, quinolizine, cinnoline, phthalazine, quinazoline, andquinoxaline. In some embodiments, the substituents are halo, hydroxy,cyano, O-C₁₋₆-alkyl, C₁₋₆-alkyl, hydroxy-C₁₋₆-alkyl, andamino-C₁₋₆-alkyl.

As used herein, the term “guanidinium group” refers to a moiety havingthe formula:

where R¹, R², R³, and R⁴ are as described above and the wavy linerepresent the point of attachment of the group to the rest of amolecule.Methods of Preparation

The compounds disclosed herein may be synthesized by methods describedbelow, or by modification of these methods. Ways of modifying themethodology include, among others, temperature, solvent, reagents etc.,and will be obvious to those skilled in the art. In general, during anyof the processes for preparation of the compounds disclosed herein, itmay be necessary and/or desirable to protect sensitive or reactivegroups on any of the molecules concerned. This may be achieved by meansof conventional protecting groups, such as those described in ProtectiveGroups in Organic Chemistry (ed. J. F. W. McOmie, Plenum Press, 1973);and Greene & Wuts, Protective Groups in Organic Synthesis, John Wiley &Sons, 1991, which are both hereby incorporated herein by reference intheir entirety. The protecting groups may be removed at a convenientsubsequent stage using methods known from the art. Synthetic chemistrytransformations useful in synthesizing applicable compounds are known inthe art and include e.g. those described in R. Larock, ComprehensiveOrganic Transformations, VCH Publishers, 1989, or L. Paquette, ed.,Encyclopedia of Reagents for Organic Synthesis, John Wiley and Sons,1995, which are both hereby incorporated herein by reference in theirentirety.

The transporter molecules described herein may be prepared by linkingtogether guanidinium group containing monomers and spacer unit monomers.Spacer unit monomers may be synthesized by techniques known in the art.In some embodiments, spacer unit monomers containing amine andcarboxylic terminal groups may be synthesized using techniques known inthe art. For example, N-(2-aminoethyl)glycine and related spacer unitmonomers may be synthesized by coupling of a diamine compound with ahalogenated t-butyl ester as described further in Stephen A. Thomson, etal, Tetrahedron Vol. 51, No. 22, pp. 6179-6194, 1995, which isincorporated herein by reference in its entirety. The t-butyl group maythen be removed by treatment with an acid. In some embodiments, the2-amino group may be functionalized, such as with a purine or pyrimidinegroup, by first protecting the terminal amine group, such as with Fmoc,and then reacting with the carboxylic functionalized purine orpyrimidine group, as described further in Stephen A. Thomson, et al,Tetrahedron Vol. 51, No. 22, pp. 6179-6194, 1995.

The guanidinium group containing monomers may be synthesized bytechniques known in the art. In some embodiments, the guanidinium groupcontaining monomers are arginine residues or derivatives thereof.

Formation of polymers containing the guanidinium and spacer unitmonomers may be obtained by using commercially available solid phasetechniques. For example, the polymer may be grown on a Rink Amide MBHAresin. Fmoc protected guanidinium containing peptides may be coupled inthe presence of PyBOP/DIPEA/DMAP and subsequently deprotected bytreatment with Ac₂O/DIPEA/DMF and 20% Piperidine/DMF. Fmoc protectedspacer unit monomers, such as peptide nucleic acid monomers, may besimilarly coupled in the presence of PyBOP/DIPEA/DMAP. These couplingsteps may be repeated in any desired order to obtain the desiredpolymer.

Although the polymer linkages in the method described above involveformation of peptide bonds, those of skill in the art will recognizeother functional groups that could be used to form linkages between themonomer units of the molecule, such as by forming ester bonds betweenthe monomers.

Where the processes for the preparation of the compounds disclosedherein give rise to mixtures of stereoisomers, such isomers may beseparated by conventional techniques such as preparative chiralchromatography. The compounds may be prepared in racemic form orindividual enantiomers may be prepared by stereoselective synthesis orby resolution. The compounds may be resolved into their componentenantiomers by standard techniques, such as the formation ofdiastereomeric pairs by salt formation with an optically active acid,such as (−)-di-p-toluoyl-d-tartaric acid and/or(+)-di-p-toluoyl-1-tartaric acid, followed by fractional crystallizationand regeneration of the free base. The compounds may also be resolvedusing a chiral auxiliary by formation of diastereomeric derivatives suchas esters, amides or ketals followed by chromatographic separation andremoval of the chiral auxiliary.

Methods of Use

In some embodiments, the transporter molecules described above may beused to enhance transport of a drug across a cell membrane. In oneembodiment, enhancing transport of a drug across a cell membranecomprises linking a therapeutic moiety to the transporter molecule. Thetherapeutic moiety may be any suitable radical of a therapeuticallyeffective drug. In some embodiments, the therapeutic moiety is linked tothe transporter molecule via a cleavable linker such that when themolecule is within a cell, the therapeutic moiety is cleaved, generatingthe drug molecule. In some embodiments, a targeting moiety is alsoattached to the transporter molecule. The targeting moiety may be amoiety that preferentially binds to receptors on one or more cell types.Thus, the transporter molecule will have increased affinity toparticular cell types, and thus increased transport across the membraneof those cell types. In this way, a drug can be effectively deliveredspecifically to the interior of cells where the drug will have its mostbeneficial effect.

In some embodiments, the cell membrane across which the transportermolecule passes is a eukaryotic cell membrane, such as the membrane inmammalian cells, cancer cells, insect cells, plant cells, or yeastcells. In other embodiments, the membrane is a prokaryotic cellmembrane, such as bacteria cell membrane.

In some embodiments, a method is provided for converting a drug to aform that is more bioavailable. The method may comprise linking the drugto a transporter molecule as described herein. The transporter moleculewill exhibit enhanced transport across cellular membranes and thus theattached drug will be more bioavailable than when delivered alone.

In some embodiments, the transporter molecules described herein maysimilarly be used to enhance transport of a drug across epithelialtissue, such as across skin, mucosmembrane, and brain blood barriers.

Pharmaceutical Compositions

In another aspect, the present disclosure relates to a pharmaceuticalcomposition comprising physiologically acceptable surface active agents,carriers, diluents, excipients, smoothing agents, suspension agents,film forming substances, and coating assistants, or a combinationthereof; and a compound disclosed herein. Acceptable carriers ordiluents for therapeutic use are well known in the pharmaceutical art,and are described, for example, in Remington's Pharmaceutical Sciences,18th Ed., Mack Publishing Co., Easton, Pa. (1990), which is incorporatedherein by reference in its entirety. Preservatives, stabilizers, dyes,sweeteners, fragrances, flavoring agents, and the like may be providedin the pharmaceutical composition. For example, sodium benzoate,ascorbic acid and esters of p-hydroxybenzoic acid may be added aspreservatives. In addition, antioxidants and suspending agents may beused. In various embodiments, alcohols, esters, sulfated aliphaticalcohols, and the like may be used as surface active agents; sucrose,glucose, lactose, starch, crystallized cellulose, mannitol, lightanhydrous silicate, magnesium aluminate, magnesium methasilicatealuminate, synthetic aluminum silicate, calcium carbonate, sodium acidcarbonate, calcium hydrogen phosphate, calcium carboxymethyl cellulose,and the like may be used as excipients; magnesium stearate, talc,hardened oil and the like may be used as smoothing agents; coconut oil,olive oil, sesame oil, peanut oil, soya may be used as suspension agentsor lubricants; cellulose acetate phthalate as a derivative of acarbohydrate such as cellulose or sugar, or methylacetate-methacrylatecopolymer as a derivative of polyvinyl may be used as suspension agents;and plasticizers such as ester phthalates and the like may be used assuspension agents.

The term “pharmaceutical composition” refers to a mixture of a compounddisclosed herein with other chemical components, such as diluents orcarriers. The pharmaceutical composition facilitates administration ofthe compound to an organism. Multiple techniques of administering acompound exist in the art including, but not limited to, oral,injection, aerosol, parenteral, and topical administration.Pharmaceutical compositions can also be obtained by reacting compoundswith inorganic or organic acids such as hydrochloric acid, hydrobromicacid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid,ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and thelike.

The term “carrier” defines a chemical compound that facilitates theincorporation of a compound into cells or tissues. For example dimethylsulfoxide (DMSO) is a commonly utilized carrier as it facilitates theuptake of many organic compounds into the cells or tissues of anorganism.

The term “diluent” defines chemical compounds diluted in water that willdissolve the compound of interest as well as stabilize the biologicallyactive form of the compound. Salts dissolved in buffered solutions areutilized as diluents in the art. One commonly used buffered solution isphosphate buffered saline because it mimics the salt conditions of humanblood. Since buffer salts can control the pH of a solution at lowconcentrations, a buffered diluent rarely modifies the biologicalactivity of a compound.

The term “physiologically acceptable” defines a carrier or diluent thatdoes not abrogate the biological activity and properties of thecompound.

The pharmaceutical compositions described herein can be administered toa human patient per se, or in pharmaceutical compositions where they aremixed with other active ingredients, as in combination therapy, orsuitable carriers or excipient(s). Techniques for formulation andadministration of the compounds of the instant application may be foundin “Remington's Pharmaceutical Sciences,” Mack Publishing Co., Easton,Pa., 18th edition, 1990.

Suitable routes of administration may, for example, include oral,rectal, transmucosal, topical, or intestinal administration; parenteraldelivery, including intramuscular, subcutaneous, intravenous,intramedullary injections, as well as intrathecal, directintraventricular, intraperitoneal, intranasal, or intraocularinjections. The compounds can also be administered in sustained orcontrolled release dosage forms, including depot injections, osmoticpumps, pills, transdermal (including electrotransport) patches, and thelike, for prolonged and/or timed, pulsed administration at apredetermined rate.

The pharmaceutical compositions of the present invention may bemanufactured in a manner that is itself known, e.g., by means ofconventional mixing, dissolving, granulating, dragee-making, levigating,emulsifying, encapsulating, entrapping or tabletting processes.

Pharmaceutical compositions for use in accordance with the presentinvention thus may be formulated in conventional manner using one ormore physiologically acceptable carriers comprising excipients andauxiliaries which facilitate processing of the active compounds intopreparations which can be used pharmaceutically. Proper formulation isdependent upon the route of administration chosen. Any of the well-knowntechniques, carriers, and excipients may be used as suitable and asunderstood in the art; e.g., in Remington's Pharmaceutical Sciences,above.

Injectables can be prepared in conventional forms, either as liquidsolutions or suspensions, solid forms suitable for solution orsuspension in liquid prior to injection, or as emulsions. Suitableexcipients are, for example, water, saline, dextrose, mannitol, lactose,lecithin, albumin, sodium glutamate, cysteine hydrochloride, and thelike. In addition, if desired, the injectable pharmaceuticalcompositions may contain minor amounts of nontoxic auxiliary substances,such as wetting agents, pH buffering agents, and the like.Physiologically compatible buffers include, but are not limited to,Hanks's solution, Ringer's solution, or physiological saline buffer. Ifdesired, absorption enhancing preparations (for example, liposomes), maybe utilized.

For transmucosal administration, penetrants appropriate to the barrierto be permeated may be used in the formulation.

Pharmaceutical formulations for parenteral administration, e.g., bybolus injection or continuous infusion, include aqueous solutions of theactive compounds in water-soluble form. Additionally, suspensions of theactive compounds may be prepared as appropriate oily injectionsuspensions. Suitable lipophilic solvents or vehicles include fatty oilssuch as sesame oil, or other organic oils such as soybean, grapefruit oralmond oils, or synthetic fatty acid esters, such as ethyl oleate ortriglycerides, or liposomes. Aqueous injection suspensions may containsubstances which increase the viscosity of the suspension, such assodium carboxymethyl cellulose, sorbitol, or dextran. Optionally, thesuspension may also contain suitable stabilizers or agents that increasethe solubility of the compounds to allow for the preparation of highlyconcentrated solutions. Formulations for injection may be presented inunit dosage form, e.g., in ampoules or in multi-dose containers, with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. Alternatively, the active ingredient may be in powder form forconstitution with a suitable vehicle, e.g., sterile pyrogen-free water,before use.

For oral administration, the compounds can be formulated readily bycombining the active compounds with pharmaceutically acceptable carrierswell known in the art. Such carriers enable the compounds of theinvention to be formulated as tablets, pills, dragees, capsules,liquids, gels, syrups, slurries, suspensions and the like, for oralingestion by a patient to be treated. Pharmaceutical preparations fororal use can be obtained by combining the active compounds with solidexcipient, optionally grinding a resulting mixture, and processing themixture of granules, after adding suitable auxiliaries, if desired, toobtain tablets or dragee cores. Suitable excipients are, in particular,fillers such as sugars, including lactose, sucrose, mannitol, orsorbitol; cellulose preparations such as, for example, maize starch,wheat starch, rice starch, potato starch, gelatin, gum tragacanth,methyl cellulose, hydroxypropylmethyl-cellulose, sodiumcarboxymethylcellulose, and/or polyvinylpyrrolidone (PVP). If desired,disintegrating agents may be added, such as the cross-linked polyvinylpyrrolidone, agar, or alginic acid or a salt thereof such as sodiumalginate. Dragee cores are provided with suitable coatings. For thispurpose, concentrated sugar solutions may be used, which may optionallycontain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel,polyethylene glycol, and/or titanium dioxide, lacquer solutions, andsuitable organic solvents or solvent mixtures. Dyestuffs or pigments maybe added to the tablets or dragee coatings for identification or tocharacterize different combinations of active compound doses. For thispurpose, concentrated sugar solutions may be used, which may optionallycontain gum arabic, talc, polyvinyl pyrrolidone, carbopol gel,polyethylene glycol, and/or titanium dioxide, lacquer solutions, andsuitable organic solvents or solvent mixtures. Dyestuffs or pigments maybe added to the tablets or dragee coatings for identification or tocharacterize different combinations of active compound doses.

Pharmaceutical preparations which can be used orally include push-fitcapsules made of gelatin, as well as soft, sealed capsules made ofgelatin and a plasticizer, such as glycerol or sorbitol. The push-fitcapsules can contain the active ingredients in admixture with fillersuch as lactose, binders such as starches, and/or lubricants such astalc or magnesium stearate and, optionally, stabilizers. In softcapsules, the active compounds may be dissolved or suspended in suitableliquids, such as fatty oils, liquid paraffin, or liquid polyethyleneglycols. In addition, stabilizers may be added. All formulations fororal administration should be in dosages suitable for suchadministration.

For buccal administration, the compositions may take the form of tabletsor lozenges formulated in conventional manner.

For administration by inhalation, the compounds for use according to thepresent invention are conveniently delivered in the form of an aerosolspray presentation from pressurized packs or a nebulizer, with the useof a suitable propellant, e.g., dichlorodifluoromethane,trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide orother suitable gas. In the case of a pressurized aerosol the dosage unitmay be determined by providing a valve to deliver a metered amount.Capsules and cartridges of, e.g., gelatin for use in an inhaler orinsufflator may be formulated containing a powder mix of the compoundand a suitable powder base such as lactose or starch.

Further disclosed herein are various pharmaceutical compositions wellknown in the pharmaceutical art for uses that include intraocular,intranasal, and intraauricular delivery. Suitable penetrants for theseuses are generally known in the art. Pharmaceutical compositions forintraocular delivery include aqueous ophthalmic solutions of the activecompounds in water-soluble form, such as eyedrops, or in gellan gum(Shedden et al., Clin. Ther., 23(3):440-50 (2001)) or hydrogels (Mayeret al., Ophthalmologica, 210(2):101-3 (1996)); ophthalmic ointments;ophthalmic suspensions, such as microparticulates, drug-containing smallpolymeric particles that are suspended in a liquid carrier medium(Joshi, A., J. Ocul. Pharmacol., 10(1):29-45 (1994)), lipid-solubleformulations (Alm et al., Prog. Clin. Biol. Res., 312:447-58 (1989)),and microspheres (Mordenti, Toxicol. Sci., 52(1):101-6 (1999)); andocular inserts. All of the above-mentioned references, are incorporatedherein by reference in their entireties. Such suitable pharmaceuticalformulations are most often and preferably formulated to be sterile,isotonic and buffered for stability and comfort. Pharmaceuticalcompositions for intranasal delivery may also include drops and spraysoften prepared to simulate in many respects nasal secretions to ensuremaintenance of normal ciliary action. As disclosed in Remington'sPharmaceutical Sciences, 18th Ed., Mack Publishing Co., Easton, Pa.(1990), which is incorporated herein by reference in its entirety, andwell-known to those skilled in the art, suitable formulations are mostoften and preferably isotonic, slightly buffered to maintain a pH of 5.5to 6.5, and most often and preferably include antimicrobialpreservatives and appropriate drug stabilizers. Pharmaceuticalformulations for intraauricular delivery include suspensions andointments for topical application in the ear. Common solvents for suchaural formulations include glycerin and water.

The compounds may also be formulated in rectal compositions such assuppositories or retention enemas, e.g., containing conventionalsuppository bases such as cocoa butter or other glycerides.

In addition to the formulations described previously, the compounds mayalso be formulated as a depot preparation. Such long acting formulationsmay be administered by implantation (for example subcutaneously orintramuscularly) or by intramuscular injection. Thus, for example, thecompounds may be formulated with suitable polymeric or hydrophobicmaterials (for example as an emulsion in an acceptable oil) or ionexchange resins, or as sparingly soluble derivatives, for example, as asparingly soluble salt.

For hydrophobic compounds, a suitable pharmaceutical carrier may be acosolvent system comprising benzyl alcohol, a nonpolar surfactant, awater-miscible organic polymer, and an aqueous phase. A common cosolventsystem used is the VPD co-solvent system, which is a solution of 3% w/vbenzyl alcohol, 8% w/v of the nonpolar surfactant Polysorbate 80™, and65% w/v polyethylene glycol 300, made up to volume in absolute ethanol.Naturally, the proportions of a co-solvent system may be variedconsiderably without destroying its solubility and toxicitycharacteristics. Furthermore, the identity of the co-solvent componentsmay be varied: for example, other low-toxicity nonpolar surfactants maybe used instead of POLYSORBATE 80™; the fraction size of polyethyleneglycol may be varied; other biocompatible polymers may replacepolyethylene glycol, e.g., polyvinyl pyrrolidone; and other sugars orpolysaccharides may substitute for dextrose.

Alternatively, other delivery systems for hydrophobic pharmaceuticalcompounds may be employed. Liposomes and emulsions are well knownexamples of delivery vehicles or carriers for hydrophobic drugs. Certainorganic solvents such as dimethylsulfoxide also may be employed,although usually at the cost of greater toxicity. Additionally, thecompounds may be delivered using a sustained-release system, such assemipermeable matrices of solid hydrophobic polymers containing thetherapeutic agent. Various sustained-release materials have beenestablished and are well known by those skilled in the art.Sustained-release capsules may, depending on their chemical nature,release the compounds for a few weeks up to over 100 days. Depending onthe chemical nature and the biological stability of the therapeuticreagent, additional strategies for protein stabilization may beemployed.

Additional therapeutic or diagnostic agents may be incorporated into thepharmaceutical compositions. Alternatively or additionally,pharmaceutical compositions may be combined with other compositions thatcontain other therapeutic or diagnostic agents.

Methods of Administration

The compounds or pharmaceutical compositions may be administered to thepatient by any suitable means. Non-limiting examples of methods ofadministration include, among others, (a) administration though oralpathways, which administration includes administration in capsule,tablet, granule, spray, syrup, or other such forms; (b) administrationthrough non-oral pathways such as rectal, vaginal, intraurethral,intraocular, intranasal, or intraauricular, which administrationincludes administration as an aqueous suspension, an oily preparation orthe like or as a drip, spray, suppository, salve, ointment or the like;(c) administration via injection, subcutaneously, intraperitoneally,intravenously, intramuscularly, intradermally, intraorbitally,intracapsularly, intraspinally, intrasternally, or the like, includinginfusion pump delivery; (d) administration locally such as by injectiondirectly in the renal or cardiac area, e.g., by depot implantation; aswell as (e) administration topically; as deemed appropriate by those ofskill in the art for bringing the compound of the invention into contactwith living tissue.

Pharmaceutical compositions suitable for administration includecompositions where the active ingredients are contained in an amounteffective to achieve its intended purpose. The therapeutically effectiveamount of the compounds disclosed herein required as a dose will dependon the therapeutic moiety incorporated, the route of administration, thetype of animal, including human, being treated, and the physicalcharacteristics of the specific animal under consideration. The dose canbe tailored to achieve a desired effect, but will depend on such factorsas weight, diet, concurrent medication and other factors which thoseskilled in the medical arts will recognize. More specifically, atherapeutically effective amount means an amount of compound effectiveto prevent, alleviate or ameliorate symptoms of disease or prolong thesurvival of the subject being treated. Determination of atherapeutically effective amount is well within the capability of thoseskilled in the art, especially in light of the detailed disclosureprovided herein.

As will be readily apparent to one skilled in the art, the useful invivo dosage to be administered and the particular mode of administrationwill vary depending upon the age, weight and mammalian species treated,the particular compounds employed, and the specific use for which thesecompounds are employed. The determination of effective dosage levels,that is the dosage levels necessary to achieve the desired result, canbe accomplished by one skilled in the art using routine pharmacologicalmethods. Typically, human clinical applications of products arecommenced at lower dosage levels, with dosage level being increaseduntil the desired effect is achieved. Alternatively, acceptable in vitrostudies can be used to establish useful doses and routes ofadministration of the compositions identified by the present methodsusing established pharmacological methods.

In non-human animal studies, applications of potential products arecommenced at higher dosage levels, with dosage being decreased until thedesired effect is no longer achieved or adverse side effects disappear.The dosage may range broadly, depending upon the desired affects and thetherapeutic indication. Typically, dosages may be between about 10microgram/kg and 100 mg/kg body weight, preferably between about 100microgram/kg and 10 mg/kg body weight. Alternatively dosages may bebased and calculated upon the surface area of the patient, as understoodby those of skill in the art.

The exact formulation, route of administration and dosage for thepharmaceutical compositions of the present invention can be chosen bythe individual physician in view of the patient's condition. (See e.g.,Fingl et al. 1975, in “The Pharmacological Basis of Therapeutics”, whichis hereby incorporated herein by reference in its entirety, withparticular reference to Ch. 1, p. 1). Typically, the dose range of thecomposition administered to the patient can be from about 0.5 to 1000mg/kg of the patient's body weight. The dosage may be a single one or aseries of two or more given in the course of one or more days, as isneeded by the patient. In instances where human dosages for compoundshave been established for at least some condition, the present inventionwill use those same dosages, or dosages that are between about 0.1% and500%, more preferably between about 25% and 250% of the establishedhuman dosage. Where no human dosage is established, as will be the casefor newly-discovered pharmaceutical compounds, a suitable human dosagecan be inferred from ED₅₀ or ID₅₀ values, or other appropriate valuesderived from in vitro or in vivo studies, as qualified by toxicitystudies and efficacy studies in animals.

It should be noted that the attending physician would know how to andwhen to terminate, interrupt, or adjust administration due to toxicityor organ dysfunctions. Conversely, the attending physician would alsoknow to adjust treatment to higher levels if the clinical response werenot adequate (precluding toxicity). The magnitude of an administrateddose in the management of the disorder of interest will vary with theseverity of the condition to be treated and to the route ofadministration. The severity of the condition may, for example, beevaluated, in part, by standard prognostic evaluation methods. Further,the dose and perhaps dose frequency, will also vary according to theage, body weight, and response of the individual patient. A programcomparable to that discussed above may be used in veterinary medicine.

Although the exact dosage will be determined on a drug-by-drug basis, inmost cases, some generalizations regarding the dosage can be made. Thedaily dosage regimen for an adult human patient may be, for example, anoral dose of between 0.1 mg and 2000 mg of each active ingredient,preferably between 1 mg and 500 mg, e.g. 5 to 200 mg. In otherembodiments, an intravenous, subcutaneous, or intramuscular dose of eachactive ingredient of between 0.01 mg and 100 mg, preferably between 0.1mg and 60 mg, e.g. 1 to 40 mg is used. In cases of administration of apharmaceutically acceptable salt, dosages may be calculated as the freebase. In some embodiments, the composition is administered 1 to 4 timesper day. Alternatively the compositions of the invention may beadministered by continuous intravenous infusion, preferably at a dose ofeach active ingredient up to 1000 mg per day. As will be understood bythose of skill in the art, in certain situations it may be necessary toadminister the compounds disclosed herein in amounts that exceed, oreven far exceed, the above-stated, preferred dosage range in order toeffectively and aggressively treat particularly aggressive diseases orinfections. In some embodiments, the compounds will be administered fora period of continuous therapy, for example for a week or more, or formonths or years.

Dosage amount and interval may be adjusted individually to provideplasma levels of the active moiety which are sufficient to maintain themodulating effects, or minimal effective concentration (MEC). The MECwill vary for each compound but can be estimated from in vitro data.Dosages necessary to achieve the MEC will depend on individualcharacteristics and route of administration. However, HPLC assays orbioassays can be used to determine plasma concentrations.

Dosage intervals can also be determined using MEC value. Compositionsshould be administered using a regimen which maintains plasma levelsabove the MEC for 10-90% of the time, preferably between 30-90% and mostpreferably between 50-90%.

In cases of local administration or selective uptake, the effectivelocal concentration of the drug may not be related to plasmaconcentration.

The amount of composition administered will, of course, be dependent onthe subject being treated, on the subject's weight, the severity of theaffliction, the manner of administration and the judgment of theprescribing physician.

Compounds disclosed herein can be evaluated for efficacy and toxicityusing known methods. For example, the toxicology of a particularcompound, or of a subset of the compounds, sharing certain chemicalmoieties, may be established by determining in vitro toxicity towards acell line, such as a mammalian, and preferably human, cell line. Theresults of such studies are often predictive of toxicity in animals,such as mammals, or more specifically, humans. Alternatively, thetoxicity of particular compounds in an animal model, such as mice, rats,rabbits, or monkeys, may be determined using known methods. The efficacyof a particular compound may be established using several recognizedmethods, such as in vitro methods, animal models, or human clinicaltrials. Recognized in vitro models exist for nearly every class ofcondition, including but not limited to cancer, cardiovascular disease,and various immune dysfunction. Similarly, acceptable animal models maybe used to establish efficacy of chemicals to treat such conditions.When selecting a model to determine efficacy, the skilled artisan can beguided by the state of the art to choose an appropriate model, dose, androute of administration, and regime. Of course, human clinical trialscan also be used to determine the efficacy of a compound in humans.

The compositions may, if desired, be presented in a pack or dispenserdevice which may contain one or more unit dosage forms containing theactive ingredient. The pack may for example comprise metal or plasticfoil, such as a blister pack. The pack or dispenser device may beaccompanied by instructions for administration. The pack or dispensermay also be accompanied with a notice associated with the container inform prescribed by a governmental agency regulating the manufacture,use, or sale of pharmaceuticals, which notice is reflective of approvalby the agency of the form of the drug for human or veterinaryadministration. Such notice, for example, may be the labeling approvedby the U.S. Food and Drug Administration for prescription drugs, or theapproved product insert. Compositions comprising a compound of theinvention formulated in a compatible pharmaceutical carrier may also beprepared, placed in an appropriate container, and labeled for treatmentof an indicated condition.

EXAMPLES Example 1 Synthesis of 5-methylpyrimidine-2,4,-dione PNAMonomer

An Fmoc protected 5-methylpyrimidine-2,4,-dione peptide nucleic acidmonomer was synthesized according to the technique described in StephenA. Thomson, et al, Tetrahedron Vol. 51, No. 22, pp. 6179-6194, 1995,which is incorporated herein by reference in its entirety. The followingsynthetic scheme was conducted:

Example 2 Synthesis of PNA Transporter Molecule

A peptide nucleic acid polymer containing guanidinium side groups wassynthesized using solid phase techniques on commercially available RinkAmide MBHA resins and Fmoc protected amino acids. An arginine monomerwas attached to the resin according to the following scheme:

The PNA monomer from Example 1 was then coupled to the arginine residueby the following reaction:

The above reactions were repeated 8 more times to obtain the followingPNA polymer:

Biotin was added as a model terminal group according to the followingreaction:

Finally, the polymer was decoupled from the resin by treating withTFA-H₂O-TIS (95:2.5:2.5).

The resulting peptide was purified using a C18 reverse phase HPLC columnwith CH3CN/aq. 0.1% TFA gradient elution and the structure was confirmedby mass spectrometry.

Example 3 Transduction Assay

Hela 705 cells were seeded at 1.5×10⁴/well in 96-well plate and culturedovernight (about 60% confluenced). The cells were then rinsed twice withPBS. The compound synthesized in Example 2 was conjugated withstreptavidin containing a FITC florescent dye. 100 μM of the produce inPBS (pH: 7.4) was applied to the cell cultures and the cultures wereincubated for 30 min at 37° C. The test compound solution was thenremoved and the cells were rinsed twice with PBS. The cells were fixedby treatment with 0.2% Glutaraldehyde in PBS for 5 min at roomtemperature. The cells were again rinsed twice with PBS and thenincubated with 10% methanol in PBS for 10 min at room temperatures. Thecells were then rinsed twice with PBS and incubated with 10% FBS in DMEMfor 30 min at 37° C. Next, the cells were rinsed once with PBS andincubated with SA-FITC (1:100 diluted in PBS) for 30 min at 37° C.Finally, the cells were again rinsed twice with PBS and observed under afluorescence microscope.

FIG. 1 depicts fluorescence micrographs of cells exposed to the oligomertest compound and cells only exposed to PBS. The micrographs illustratethat the test compound was taken up into the cells.

Example 4 Transdermal Crossing

Female nude mice (NU/NU) were injected with anesthesia (Ketaset 25mg/ml, Xylazine 5 mg/ml, 50 μl/mouse). 130 μl of the compound fromExample 2 in PBS was then applied to the skin of the mice. After 1 hourof treatment, skin samples were taken and frozen with OCT compoundimmediately by liquid nitrogen. The frozen samples were sliced to 5 μmthickness and placed on glass slides.

The frozen tissue slides were dried at room temperature and fixed inacetone for 10 minutes. The samples were then rinsed in PBS twice for 10min each with gentle shaking at 40˜50 RPM. The samples werepre-incubated with a blocking reagent (0.2% gelatin in PBS) for 30 minat room temperature. The samples were then incubated with optimaldiluted substrate for 1 hr at room temperature (Streptavidin-FITC, 1:100diluted in PBS). After rinsing in PBS for 10 min with gentle shaking at40˜50 RPM, the samples were soaked in hematoxylin for 2 min. Finally,the samples were again rinsed in PBS for 10 min with gentle shaking at40˜50 RPM and extra PBS was sucked out. The samples were mounted withDAKO anti-fading mounting medium and observed under a fluorescencemicroscope. The biotinylated test compound was visualized by bindingwith the streptavidin conjugated with FITC.

FIG. 2 depicts the fluorescence micrographs of tissue samples exposed tothe test compound and control samples exposed only to PBS. Themicrographs illustrate that the test compound was taken up into thetissue samples.

1. A compound having the formula:

wherein: each L is independently selected to be a linker moietycomprising a molecular fragment having a molecular weight less thanabout 1000 or L is absent; and each T is a terminal group independentlyselected from the group consisting of hydrogen, hydroxy, an amine group,a carboxylic group, a N-terminal peptide, a C-terminal peptide, areporting moiety, a targeting moiety, and a therapeutic moiety, or eachT may be independently absent.
 2. A compound having the formula:

wherein: each L is independently selected to be a linker moietycomprising a molecular fragment having a molecular weight less thanabout 1000 or L is absent; and each T is a terminal group independentlyselected from the group consisting of hydrogen, hydroxy, an amine group,a carboxylic group, a N-terminal peptide, a C-terminal peptide, areporting moiety, a targeting moiety, and a therapeutic moiety, or eachT may be independently absent.
 3. A method for enhancing transport of abiologically active moiety across a biological membrane, comprisingcontacting a biological membrane with a compound of claim 1 or claim 2,wherein at least one T comprises a moiety selected from the groupconsisting of a reporting moiety, a targeting moiety, and a therapeuticmoiety, whereby said contacting is effective to promote transport ofsaid compound across said biological membrane at a rate that is greaterthan a trans-membrane transport rate of the moiety in non-conjugatedform.
 4. The method of claim 3, wherein said biological membrane is aeukaryotic cell membrane.
 5. The method of claim 4, wherein theeukaryotic cells selected from the group consisting of mammalian cells,cancer cells, insect cells, plant cells, and yeast cells.
 6. The methodof claim 3, wherein said biological membrane is a prokaryotic cellmembrane.
 7. The method of claim 3, wherein said biological membrane isa cancer cell membrane.
 8. The method of claim 3, wherein at least one Tcomprises a therapeutic moiety.
 9. The method of claim 8, wherein saidtherapeutic moiety is an anti-cancer agent.
 10. A pharmaceuticalcomposition, comprising a compound of claim 1 or claim 2 and apharmaceutically acceptable carrier, diluent, or excipient.